采用新鲜分离自健康供体的外周血单个核细胞对化合物的抗炎特性进行体外检测。
In vitro testing for anti-inflammatory properties of compounds employing peripheral blood mononuclear cells freshly isolated from healthy donors.
机构信息
Division of Biological Chemistry, Biocenter, Innsbruck Medical University, Fritz Pregl Strasse 3, 6020 Innsbruck, Austria.
出版信息
Inflamm Res. 2011 Feb;60(2):127-35. doi: 10.1007/s00011-010-0244-y. Epub 2010 Aug 26.
INTRODUCTION
Inflammation is crucially involved in a variety of diseases like autoimmune syndromes, cardiovascular and neurodegenerative disorders, cancer, sepsis and allograft rejection.
METHODS
Freshly isolated human peripheral blood mononuclear cells (PBMCs) are used as a screening assay for anti-inflammatory properties of compounds. Determinations of neopterin production by ELISA and of tryptophan degradation by HPLC are used as read-outs. Results are compared with further markers of immune response and oxidative stress.
RESULTS
Phytohaemagglutinin induced significant tryptophan degradation and neopterin formation in PBMC, which correlated with IFN-γ, TNF-α, soluble cytokine receptors and isoprostane-8. Addition of vitamin C and E suppressed the responses dose-dependently.
DISCUSSION
The determination of tryptophan degradation and neopterin production in PBMC reflects various pro- and anti-inflammatory cascades that are of relevance also in patients. It constitutes a robust and reliable approach to screen anti-inflammatory or immunosuppressive drugs and may improve throughput, speed and cost-effectiveness in drug discovery.
简介
炎症在多种疾病中起着关键作用，如自身免疫综合征、心血管和神经退行性疾病、癌症、败血症和同种异体移植排斥。
方法
新鲜分离的人外周血单核细胞 (PBMC) 被用作化合物抗炎特性的筛选检测。通过 ELISA 测定新蝶呤的产生和 HPLC 测定色氨酸的降解作为读出。结果与免疫反应和氧化应激的进一步标志物进行比较。
结果
植物血球凝集素诱导 PBMC 中显著的色氨酸降解和新蝶呤形成，这与 IFN-γ、TNF-α、可溶性细胞因子受体和异前列烷-8 相关。维生素 C 和 E 的添加以剂量依赖性方式抑制反应。
讨论
PBMC 中色氨酸降解和新蝶呤产生的测定反映了各种促炎和抗炎级联反应，这些反应在患者中也很重要。它构成了一种强大而可靠的筛选抗炎或免疫抑制药物的方法，并可能提高药物发现的通量、速度和成本效益。
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